EP1050578B1 - Appareil et procédés de culture de materiau biologique - Google Patents

Appareil et procédés de culture de materiau biologique Download PDF

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Publication number
EP1050578B1
EP1050578B1 EP00113775A EP00113775A EP1050578B1 EP 1050578 B1 EP1050578 B1 EP 1050578B1 EP 00113775 A EP00113775 A EP 00113775A EP 00113775 A EP00113775 A EP 00113775A EP 1050578 B1 EP1050578 B1 EP 1050578B1
Authority
EP
European Patent Office
Prior art keywords
micro
organic polymer
layer
culturing
coating
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Expired - Lifetime
Application number
EP00113775A
Other languages
German (de)
English (en)
Other versions
EP1050578A1 (fr
Inventor
Jess Paul Fuller
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Ashby Scientific Ltd
Original Assignee
Ashby Scientific Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Ashby Scientific Ltd filed Critical Ashby Scientific Ltd
Priority claimed from EP96928558A external-priority patent/EP0847441B1/fr
Publication of EP1050578A1 publication Critical patent/EP1050578A1/fr
Application granted granted Critical
Publication of EP1050578B1 publication Critical patent/EP1050578B1/fr
Anticipated expiration legal-status Critical
Expired - Lifetime legal-status Critical Current

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M27/00—Means for mixing, agitating or circulating fluids in the vessel
    • C12M27/10—Rotating vessel
    • C12M27/12—Roller bottles; Roller tubes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/20—Material Coatings
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
    • C12M25/06—Plates; Walls; Drawers; Multilayer plates

Definitions

  • the present invention relates to vessels having textured surfaces for growing animal cells, plant cells, micro-organisms and the like ('bio-substances') and to methods for their production.
  • bio-substances can be propagated in vitro in containers such as so-called 'roller bottles', that is, cylindrical vessels, usually of glass or polystyrene, which are partially filled with a liquid medium comprising the bio-substances.
  • the bottles are arranged to be rotated slowly about their longitudinal axes, promoting aeration, while the liquid medium provides nutrients to the bio-substances as they grow on the inner surfaces of the container.
  • apparatus for use in a method of culturing micro-biological material, wherein said apparatus has a rough or uneven growth surface with a plurality of craters or crater-like depressions defined in the inner surface of the apparatus per se or in a coating of an organic polymer formed on the inner surface of the apparatus and, arranged for contact with micro-biological material being cultured.
  • the growth surface is rendered rough or uneven by a plurality of craters or crater like depressions, preferably to depths of 1mm or less and more preferably to depths of 0.1-0.5 mm.
  • the craters or crater like depressions measure less than 2mm, and more preferably less than 1mm across. Most preferably, the crater or crater like depressions measure less than 0.5mm across.
  • the growth surface is micro-cupellated.
  • the organic polymer coated on the inner surface of the apparatus is preferably a silicone rubber.
  • the invention provides a method of culturing a micro-biological material, comprising culturing said material using apparatus in accordance with its first aspect.
  • the inventive method allows biological material to be grown at an enhanced rate.
  • the invention provides a method of providing an apparatus with a rough or uneven growth surface for culturing microbiological material comprising the steps of forming a mobile surface studded with a granular material on a surface of the apparatus, setting the mobile surface and removing the granular material to leave a rough or uneven surface.
  • an article having a textured surface that can be obtained by a method according to the invention in its third aspect.
  • the article is a tissue or cell culture vessel, such as a roller-bottle.
  • the biological material preferably comprises tissue, cells or like matter, including viral particles or virions.
  • the mobile surface is preferably tacky prior to the setting step, so as to promote adherence of the granular material.
  • a (preferably solvent-less) silicone paint designed for polymerisation to form silicone rubber, is applied to the interior surface of a glass or polystyrene roller bottle. While the coating is still in liquid form, common salt particles are injected into the container, for example, under pressure, to adhere to the prepared silicone coating, with any excess salt being removed again from the container. The coating is then allowed to polymerise into a solid layer at ambient or elevated temperature, depending on the type of silicone used.
  • the salt particles adhering to the coating are removed by dissolution in water, to leave a surface-porous layer exhibiting a cratered or micro-cupellated structure, as well as the special affinity for bio-substances of silicone rubber.
  • the container so prepared is found to result in a greatly increased yield when used for growth processes in the manner described.
  • a conventional adhesive preferably a mineral spirit based primer, can be first applied to the surface prior to deposition of the silicone layer.
  • the growth surface should be non-toxic and inert in respect of bio-substances and liquid media.
  • the method of creating such surfaces should not involve any materials, which if remaining on the surface as impurities, could have a deleterious effect on the growth process.
  • various types of particles can be used in place of common salt, such as, for example, sugar, saltpetre and the like; however, common salt, sucrose or potassium nitrate are preferred as being non-toxic and inert in respect of normal media and bio-substances, as well as being soluble in water for subsequent removal. Solubility of the particles in a non-toxic solvent is an important factor in their choice.
  • surface pitting or cratering for example, micro-cupels
  • salt or other suitable particles are injected, for example, pneumatically, into the container while the latter is at elevated temperature, either during its manufacture or on subsequent heating. Upon cooling, the particles are then removed as described above, leaving the interior surface of the glass or polystyrene container with a textured, cratered or micro-cupellated structure, again resulting in enhanced efficiency of the process.
  • the standard procedure of plasma treatment of the inner surface can be applied to impart a negative electrical charge to the surface, as required for the growth process.
  • Other types of particle can be used apart from salt, the latter however, being again preferred for the reasons enumerated.
  • containers which comprise flat plates, for example, of polystyrene, wetted by the liquid medium, and upon which the bio-substances are made to grow.
  • Such plates can also be provided with coatings of silicone rubber having a cratered or micro-cupellated, porous, surface structure according to the invention, in the manner described above, to result in substantially enhanced productivity.
  • roller bottles and other containers according to the invention can have a specially textured, cratered or micro-cupellated growth surface, usually an inner surface, suitable for use in the growth of bio-substances as aforesaid.
  • these surfaces are more conducive to the adhesion and growth of bio-substances, with resulting improved process efficiency. It is considered that the larger surface areas provided by such growth surfaces is a factor in their enhanced performance.
  • a polystyrene roller bottle 1 is provided with a cratered or micro-cupellated surface according to the inventive method as follows.
  • a coating 2 of silicone rubber primer SS 4155 (General Electric Co., Connecticut, U.S.A.) is applied by brushing or other means and allowed to dry at 70°C for 15 minutes.
  • a coating 3 of silicone rubber paint RTV 118 (General Electric Co., Connecticut, U.S.A.), approximately 0.05 to 0.2mm in thickness, is applied to primer coating 2 and left to stand for 1 minute.
  • a substantial quantity of common salt 4 is then introduced into bottle 1 until approximately 1/3 full and the latter shaken, so that salt 4 adheres to all of the paint coating 3.
  • Bottle 1 is then filled with boiling water and allowed to stand for 10 minutes before the water is removed. This last step is repeated once before bottle 1 is rinsed with isopropyl alcohol and left to dry at 70°C for 30 mins.
  • the resultant roller bottle 1 has a textured interior surface 5 provided by craters or micro-cupels 6 inset in silicone rubber layer 4.
  • the roller bottle 1 can then be inoculated with a biological material, for example cells taken from a commercially available cell line such as Madin Darby Canine Kidney, suspended in an appropriate commercially available nutrient medium, such as Glucose Minimum Essential Medium containing 5% foetal calf serum, selected for its suitability for growing the chosen cells.
  • a biological material for example cells taken from a commercially available cell line such as Madin Darby Canine Kidney, suspended in an appropriate commercially available nutrient medium, such as Glucose Minimum Essential Medium containing 5% foetal calf serum, selected for its suitability for growing the chosen cells.
  • the roller bottle can then be placed upon conventional equipment designed to gently roll the bottle under conditions conducive to growing cells from the selected line. For example, a common culturing temperature is 37°C. After an appropriate culturing period, the growing cells can be harvested from the roller bottle using conventional techniques, such as trypsination or mechanical scraping.

Landscapes

  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Chemical & Material Sciences (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Sustainable Development (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Immunology (AREA)
  • Clinical Laboratory Science (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Claims (18)

  1. Appareillage destiné à être utilisé dans une méthode de culture de matériau micro biologique, dans lequel ledit appareillage comporte une surface de croissance rugueuse ou non uniforme présentant une pluralité de cratères ou de creux semblables à des cratères dans la surface interne de l'appareillage en lui-même ou dans un revêtement d'un polymère organique placé sur la surface interne de l'appareillage et destiné à venir en contact avec le matériau micro biologique cultivé.
  2. Appareillage selon la revendication 1 dans lequel le polymère organique est un caoutchouc de silicone.
  3. Appareillage selon l'une des deux revendications précédentes dans lequel les cratères ou les creux semblables à des cratères ont une profondeur de 1 mm ou moins et de préférence une profondeur de 0,1-0,5 mm.
  4. Appareillage selon l'une quelconque des revendications précédentes dans lequel les cratères ou les creux semblables à des cratères ont une dimension transversale inférieure à 2 mm, de préférence inférieure à 1 mm, et plus préférablement inférieure à 0,5 mm.
  5. Appareillage selon l'une quelconque des revendications précédentes dans lequel la surface de croissance présente des micro cupules.
  6. Une méthode pour la culture d'un matériau micro biologique consistant à cultiver ledit matériau en utilisant un appareillage selon l'une quelconque des revendications précédentes, méthode dans laquelle ledit matériau est mis en contact avec la surface de croissance rugueuse ou non uniforme.
  7. Une méthode de création d'un appareillage présentant une surface de croissance rugueuse ou non uniforme en vue de la culture d'un matériau micro biologique, méthode consistant à former une couche mobile parsemée d'une matière granulaire sur une surface de l'appareillage, à fixer la couche mobile et à éliminer la matière granulaire de manière à laisser une surface rugueuse ou non uniforme.
  8. Une méthode selon la revendication 7 dans laquelle la couche mobile est collante avant d'être fixée.
  9. Une méthode selon l'une des revendications 7 ou 8 dans laquelle la surface mobile est créée en ramollissant la matière de l'appareillage et en parsemant la matière ramollie de matière granulaire.
  10. Une méthode selon l'une des revendications 7 ou 8 dans laquelle la couche mobile est formée en revêtant la surface intérieure de l'appareillage avec une couche d'un polymère organique filmogène et en parsemant sur cette couche une matière granulaire.
  11. Une méthode selon l'une des revendications 7 ou 8 dans laquelle la couche mobile est formée en revêtant la surface interne de l'appareillage avec une couche d'un polymère organique filmogène en mélange avec une matière granulaire.
  12. Une méthode selon l'une des revendications 10 ou 11 dans laquelle le polymère organique filmogène est un matériau de type silicone constituant un caoutchouc.
  13. Une méthode selon l'une des revendications 10, 11 ou 12 dans laquelle la surface interne de l'appareillage est traitée par un adhésif avant d'être revêtue par la couche de polymère organique filmogène.
  14. Une méthode selon l'une des revendications 7-13 dans laquelle l'appareillage est réalisé en verre ou en un polymère organique.
  15. Une méthode selon l'une quelconque des revendications 7-14 dans laquelle la matière granulaire est soluble et est éliminée par dissolution.
  16. Une méthode selon la revendication 15 dans laquelle la matière granulaire est du sucrose, du sel ou du nitrate de potassium.
  17. Une méthode selon l'une quelconque des revendications 7-16 dans laquelle la surface de croissance rugueuse ou non uniforme présente des cratères ou des micro cupules.
  18. Un appareillage destiné à être utilisé pour la culture d'un matériau micro biologique et qui peut être obtenu par une méthode selon l'une quelconque des revendications 7-17.
EP00113775A 1995-08-31 1996-08-23 Appareil et procédés de culture de materiau biologique Expired - Lifetime EP1050578B1 (fr)

Applications Claiming Priority (5)

Application Number Priority Date Filing Date Title
GBGB9517717.6A GB9517717D0 (en) 1995-08-31 1995-08-31 Means for the growth of cells & micro-organisms
GB9517717 1995-08-31
GBGB9526032.9A GB9526032D0 (en) 1995-08-31 1995-12-20 Apparatus and methods for culturing biological material
GB9526032 1995-12-20
EP96928558A EP0847441B1 (fr) 1995-08-31 1996-08-23 Appareil et procedes de culture de materiau biologique

Related Parent Applications (1)

Application Number Title Priority Date Filing Date
EP96928558A Division EP0847441B1 (fr) 1995-08-31 1996-08-23 Appareil et procedes de culture de materiau biologique

Publications (2)

Publication Number Publication Date
EP1050578A1 EP1050578A1 (fr) 2000-11-08
EP1050578B1 true EP1050578B1 (fr) 2002-05-29

Family

ID=10779949

Family Applications (1)

Application Number Title Priority Date Filing Date
EP00113775A Expired - Lifetime EP1050578B1 (fr) 1995-08-31 1996-08-23 Appareil et procédés de culture de materiau biologique

Country Status (2)

Country Link
EP (1) EP1050578B1 (fr)
GB (2) GB9517717D0 (fr)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20220243021A1 (en) * 2018-08-13 2022-08-04 Sio2 Medical Products, Inc. Polymeric cell culturing surface having high cell adhesion

Family Cites Families (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB1092634A (en) * 1964-03-24 1967-11-29 Ici Ltd Improvements in or relating to supporting media for microbiological fermentation processes
JPS5187578A (ja) * 1975-01-30 1976-07-31 Matsushita Electric Works Ltd Kagakumetsukyokibanno seiho
US4320198A (en) * 1979-08-08 1982-03-16 Mynatt Roy L Fiber production from continuous cultivation of micro-organisms
US4284275A (en) * 1979-10-11 1981-08-18 Fletcher Herbert E Polyurethane gripping material
US4376826A (en) * 1980-06-17 1983-03-15 Mynatt Roy L Continuous production of bacteria for leaching of metallic ore
FR2522014A1 (fr) * 1982-02-25 1983-08-26 Pasteur Institut Support pour cultures cellulaires, ensemble de culture comportant un tel support et procede pour cultiver des cellules en presence de ce support
JPS6422934A (en) * 1987-07-16 1989-01-25 Polyplastics Co Molded plastic product having roughened surface for surface treatment and production thereof
US4859712A (en) * 1988-10-12 1989-08-22 Cox-Uphoff International Silicone foam and method for making it
US4912058A (en) * 1989-04-27 1990-03-27 Becton, Dickinson And Company Roller bottle
DE4132379A1 (de) * 1991-09-28 1993-04-08 Kernforschungsz Karlsruhe Substrat fuer zellkulturen und kultur von zellen oder zellaggregaten
US5272084A (en) * 1991-12-18 1993-12-21 Corning Incorporated Cell culture vessels having interior ridges and method for cultivating cells in same
CA2162465A1 (fr) * 1994-03-11 1995-09-14 Craig Sandstrom Bioreacteur a ecoulement continu, comportant des rainures pour la retention de cellules
DE4410718A1 (de) * 1994-03-28 1995-10-12 Bode Chemie Gmbh & Co Vorrichtung zur Keimzahlbestimmung

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Publication number Publication date
EP1050578A1 (fr) 2000-11-08
GB9517717D0 (en) 1995-11-01
GB9526032D0 (en) 1996-02-21

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